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Abstract

Viroid diseases are prevalent in all grapevine-cultivating regions around the world, affecting fruit quality and yield and posing a significant threat to global grapevine production. Meristem culture is a highly effective method for eliminating pathogens in In vitro propagated crops like grapevine. In this study, we present a protocol for producing GYSVd-2-free grapevine plantlets through apical meristem tip culture from indirect or direct organogenesis. Three different apical meristem sizes (0.5, 0.7, and 1.0 mm) were excised and cultured in Murashige and Skoog medium supplemented with different concentrations and combinations of cytokinin and auxin to determine the optimum size for viroid elimination. The RT-PCR analysis (indexing) was conducted on meristem- or callus-derived shoots of varying sizes (0.5, 0.7, and 1.0 mm). The study reveals that the 0.5 mm size was optimal for the GYSVd-2 elimination via indirect or direct organogenesis. The current research focused GYSVd-2 on detecting and eliminating in Pakistani grapevine cultivars to produce certified, healthy, viroid-free plantlets, with the potential to improve viroid-free plant production in the viticulture industry. Our investigation provides evidence that the efficiency of viroid elimination through apical meristem tip culture is an effective method for producing viroid-free grapevines in the future

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